Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse primary neuron cells labelling MAP2 with ab318993 at 1/100 (9.72 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in mouse primary neurons (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of Iba1 in a section of formalin-fixed paraffin-embedded human brain. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab318305 at 1 μg/ml, and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded human brain (positive) and human heart muscle (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cells labeling IL-2 with ab306565 at 1/100 dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (2 ug/ml) (Green). Confocal image showing cytoplasmic staining in Jurkat cells treated with TPA (40 nM) and A23187 (2 uM) for 1 h, then together with BFA (300 ng/ml) for another 23 h. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (2.5ug/ml) (Red). The nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (2 ug/ml).

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling GM130 with ab302489 at 1/1000 dilution (0.1ug) (Red) compared with a Chicken IgY (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTSD (cathepsin D) with ab302646 at 1/100 (8.1 ug/ml ) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab320709 staining KI67 in Hap1 (positive) and Hap1-KI67 (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab320709 at 1 ug/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labeled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab321866 staining ACTA2 in HeLa (positive) and A431 (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab321866 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This data was developed using the same antibody clone in a different buffer formulation.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of GFAP in sections of formalin-fixed paraffin-embedded human cerebral cortex*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (pH9.0) for 40 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of PCNA in sections of formalin-fixed paraffin-embedded human colon*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322083 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. This data was developed using the same antibody clone in a different buffer formulation. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTSD (cathepsin D) with ab302646 at 1/1000 dilution (0.1ug) (Red) compared with a Chicken IgY (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of KI67 in sections of formalin-fixed paraffin-embedded human tonsil (positive) and human brain (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab320709 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labeled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervix adenocarcinoma epithelial cell) cells labelling GM130 with ab302489 at 1/500 (1.814 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing golgi staining in HeLa cell line is observed. ab52649 Anti-GM130 rabbit monoclonal antibody - cis-Golgi Marker was used to counterstain tubulin at 1/100 5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab322083 staining PCNA in A-431 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322083 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This data was developed using the same antibody clone in a different buffer formulation.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized VIM knock out HeLa (VIM knockout human cervical adenocarcinoma epithelial cell line) cells labelling Vimentin with ab316214 at 1/5000 (0.16 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 (2ug/ml) dilution (Green).

Confocal image showing cytoplasmic staining in wild-type HeLa cell line (shown in green), and negative staining in VIM knock out HeLa cell line. The counterstain was observed in red. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 (2.5ug/ml) dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 (2ug/ml) dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Composite multiplex immunofluorescence staining of SYP, MAP2 and Tau (MBD region) staining in a section of formalin-fixed paraffin-embedded human Alzheimer’s brain*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab309493 at 1µg/ml dilution (shown in green), ab318993 at 1µg/ml (shown in magenta), and ab308439 at 1µg/ml (shown in yellow). Then incubated for 1 hour with ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed 1/1000, ab150086 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 555) preadsorbed 1/1000, and ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of VIM in sections of formalin-fixed paraffin-embedded human kidney (positive) and human skeletal muscle (negative)*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323238 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab323240 staining IL1B in wild-type THP-1 cells and IL1B knockout THP-1 cells +/- TPA (80nM overnight) and LPS (100ng/ml, 6h) cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4° C with ab323240 at 1ug/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of ACTA2 in sections of formalin-fixed paraffin-embedded human colon (positive) and human cerebellum (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour withab321866 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. This data was developed using the same antibody clone in a different buffer formulation. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling MAP2 with ab318993 at 1/100 (9.72 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : HEK-293
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab323238 staining VIM in Hap1 (positive) and Hap1-VIM KO (negative) cells. The cells were fixed with 100% methanol (5 min) permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4° C with ab323238 at 1ug/ml (shown in green) and ab202272 Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 4% paraformaldehyde (10 min).

Image was acquired with a high-content analyser (Operetta CLS Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling NEFH with ab317042 at 1/500 (2.098 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in SH-SY5Y cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control : MCF7 (PMID : 25985363).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab322912 staining LC3B in HAP1 cells (wildtype and MAP1LC3B knockout) +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322912 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab318305 staining Iba1 in THP-1 (positive) and HeLa (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab318305 at 5 µg/ml and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Composite multiplex immunofluorescence staining of Iba1, GFAP and MAP2 staining in a section of formalin-fixed paraffin-embedded human cerebral cortex*.

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with EDTA (Ph9.0) using retrieval settings of 100°C for 40 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 5µg/ml dilution (shown in green), ab300645 at 5µg/ml (shown in magenta), and ab178846 at 5µg/ml (shown in yellow). Then incubated for 1 hour with ab150119 Goat Anti-Mouse IgG H&L (Alexa Fluor® 647) preadsorbed 1/1000, ab150084 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed 1/1000, and ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed 1/1000. Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium ^®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of GFAP in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse skeletal muscle (negative).

Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab323239 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®.

Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of LC3B in sections of formalin-fixed paraffin-embedded mouse brain (positive) and mouse liver (negative). Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322912 at 1/100 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab322912 staining LC3B in NIH/3T3 +/- Chloroquine (50µM 24 hours) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322912 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling MAP2 with ab318993 at 1/1000 dilution (0.1 ug)(Red) compared with a Chicken IgY (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173) at 1/5000 dilution was used as the secondary antibody.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of PCNA in sections of formalin-fixed paraffin-embedded mouse testis. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab322083 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times. This data was developed using the same antibody clone in a different buffer formulation.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab322083 staining PCNA in NIH/3T3 cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab322083 at 1µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

This data was developed using the same antibody clone in a different buffer formulation.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling MAP2 with ab318993 at 1/1000 dilution (0.1 ug)(Red) compared with a Chicken IgY (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized rat primary neuron cells labelling MAP2 with ab318993 at 1/100 (9.72 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing cytoplasmic staining in rat primary neurons (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Confocal scanning Z step was set as 0.3 μm followed by image processing with maximum Z projection.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/500 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

ab320061 staining F4/80 in Raw264.7 (positive) and NIH/3T3 (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab320061 at 5µg/ml (shown in green) and ab202272, Alexa Fluor® 594 Rabbit monoclonal [EP1332Y] to alpha Tubulin - Microtubule Marker (shown in pseudocolour magenta). Cells were then incubated with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution. Nuclear DNA was labelled with DAPI (shown in blue). Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of ACTA2 in sections of formalin-fixed paraffin-embedded mouse stomach (positive) and mouse brain (negative)*. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab321866 at 1/500 dilution and then incubated for 1 hour with ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). This data was developed using the same antibody clone in a different buffer formulation. For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of F4/80 in sections of formalin-fixed paraffin-embedded mouse spleen (positive) and mouse pancreas (negative). Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab320061 at 1/100 dilution and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized mouse primary neural/glia cell cells labelling NEFH with ab317042 at 1/500 (2.098 ug/ml) dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing positive staining in mouse primary neuron (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab11267 Anti-MAP2 mouse monoclonal antibody was used to counterstain tubulin at 1/200 4ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab317042 at 1/50 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.

-ve control 2 : ab11267 at 1/200 dilution, followed by ab150173 Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of CD68 in a section of formalin-fixed paraffin-embedded rat spleen. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab318306 at 1/500 dilution, and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Negative Control : NIH/3T3. Flow cytometry (intracellular) analysis of NIH/3T3 (mouse embryonic fibroblasts) fixed with 4% PFA showing no staining of GFAP with ab323239 used at 1/50000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Immunofluorescence staining of Iba1 in a section of formalin-fixed paraffin-embedded mouse brain. Performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate (pH6.0) for 20 minutes. The section was then incubated at room temperature for 1 hour with ab318305 at 1 μg/ml, and then incubated for 1 hour with ab150173, Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). The section was then mounted using Dako Fluorescence Mounting Medium®. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). For other IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antigen retrieval conditions, antibody concentrations and incubation times.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Goat Anti-Chicken IgY H&L (Alexa Fluor® 488) preadsorbed (AB150173)

Flow cytometry (intracellular) analysis of Mouse primary neural/glia cells fixed with 4% PFA staining GFAP with ab323239 used at 1/50000 dilution (0.01ug/mL) (right). Isotype control Chicken IgY (left). Secondary antibody was Goat anti-Chicken IgY H&L (Alexa Fluor® 488, ab150173, used at 1/5000 dilution.